Pyrosequencing Methods and Protocols
The primary purpose of this volume is to demonstrate the range of applications of the Pyrosequencing technology in research and diagnostics and to provide detailed protocols. Beginning with an up-to-date overview of the biochemistry, the volume continues
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Ulrich Lehmann Jörg Tost Editors
Pyrosequencing Methods and Protocols Second Edition
METHODS
IN
MOLECULAR BIOLOGY
Series Editor John M. Walker School of Life and Medical Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK
For further volumes: http://www.springer.com/series/7651
Pyrosequencing Methods and Protocols
Second Edition
Edited by
Ulrich Lehmann Medizinische Hochschule Hannover, Institute of Pathology, Hannover, Germany
Jörg Tost Laboratory for Epigenetics and Environment, Centre National de Génotypage, CEA-Institut de Génomique, Evry, France
Editors Ulrich Lehmann Medizinische Hochschule Hannover Institute of Pathology Hannover, Germany
Jörg Tost Laboratory for Epigenetics and Environment Centre National de Génotypage CEA-Institut de Génomique Evry, France
ISSN 1064-3745 ISSN 1940-6029 (electronic) Methods in Molecular Biology ISBN 978-1-4939-2714-2 ISBN 978-1-4939-2715-9 (eBook) DOI 10.1007/978-1-4939-2715-9 Library of Congress Control Number: 2015939912 Springer New York Heidelberg Dordrecht London © Springer Science+Business Media New York 2015 This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, express or implied, with respect to the material contained herein or for any errors or omissions that may have been made. Printed on acid-free paper Humana Press is a brand of Springer Springer Science+Business Media LLC New York is part of Springer Science+Business Media (www.springer.com)
Preface The Pyrosequencing technology has been around for nearly 20 years and has become a widely used technology for a broad variety of applications to analyze genetic and epigenetic variation as well as to identify bacteria, viruses, and microorganisms. Pyrosequencing is a versatile, flexible, accurate, and extremely quantitative sequencing-by-synthesis technology, which is relatively easy to handle and has a short time to results. The sequence-based readout, and the rapid and standardized work-flow of Pyrosequencing as well as the improved limit of detection and analytical sensitivity compared to traditional sequencing technologies, make this technology a method of
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